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| 珍稀濒危植物裸果木RAPD反应体系优化研究 | |
| 引用本文： | 王静,王海芳,花立民,马喜迎,王亮亮.珍稀濒危植物裸果木RAPD反应体系优化研究[J].草原与草坪,2012,32(1):7-11. |
| 作者姓名： | 王静 王海芳 花立民 马喜迎 王亮亮 |
| 作者单位： | 1. 兰州职业技术学院,甘肃兰州,730070 2. 甘肃农业大学草业学院,甘肃兰州,730070 3. 甘肃省小陇山林业实验局,甘肃天水,741020 |
| 基金项目： | 公益性农业行业专项一牧区生态高效草原牧养技术模式研究与示范 |
| 摘 要： | 以裸果木新鲜叶片为材料,采用改进CTAB法提取基因组DNA,在参考一般RAPD分析反应程序的基础上,研究了裸果木RAPD分析过程中的影响因素Taq酶、Mg2+、dNTP、引物、模板DNA浓度、变性时间等,建立了适于裸果木RAPD反应的PCR体系:模板DNA为4ng/μL,引物浓度为0.3μmol/L,dNTP浓度为0.5mmol/L,Mg2+浓度为2.0mmol/L,Taq酶用量为1U,ddH2O补足25μL;94℃预变性5min,94℃变性45s,36℃退火40s,72℃延伸1min,共30个循环,最后72℃延伸10min,扩增结果稳定。 |
| 关 键 词： | 裸果木 RAPD 反应体系 优化 |
Study on optimization of rapd reaction system for endangered plant Gymnocarpos przewalskii |
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| WANG Jing,WANG Hai-fang,HUA Li-min,MA Xi-ying,WANG Liang-liang.Study on optimization of rapd reaction system for endangered plant Gymnocarpos przewalskii[J].Grassland and Turf,2012,32(1):7-11. | |
| Authors: | WANG Jing WANG Hai-fang HUA Li-min MA Xi-ying WANG Liang-liang |
| Institution: | 1.Lanzhou Vocational Technology College,Lanzhou 730070,China;2.College of Grassland Science,Gansu Agricultural University,Lanzhou 730070,China;3.Xiaolongshan Forest Experiment Bureau of Gansu Province;Tianshui 741020,China) |
| Abstract: | To study the RAPD optimal reaction system of G.przewalskii and provide the research basis for RAPD amplification.The DNA templates extracted from fresh leaves by CTAB methods,such factors impacted on RAPD analysis as Taq enzymes,Mg2+,dNTP,primer,concentration of DNA template,time of degeneration and temperature were studied,and PCR systems fitted for RAPD reaction of G.przewalskii were established.The reactions were performed in a volume of 25 μL containing 2.5μL 10×Buffer,2.5 mmol/L Mg2+,Taq DNA ploymerase 1U,0.5 mmol/L dNTPs,0.3μmol/L Primer,and 4.0 ng/μl template DNA.PCR program was 5 min at 94℃ for predenaturation,then 30 cycles of 45 s at 94 ℃ for denaturation,40 s at 36 ℃ for annealing,1 min at 72 ℃ for extension,finally extension at 72 ℃ for 10 min. |
| Keywords: | Gymnocarpos przewalskii RAPD reaction system optimization |
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